Difference between revisions of "Methods"

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* introduction of markerfree deletions: {{PubMed|24140578,23916947,23911571,22422839,20057169}}
 
* introduction of markerfree deletions: {{PubMed|24140578,23916947,23911571,22422839,20057169}}
  
* generation of markerfree mutations: {{PubMed|20134232,21542786}}
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* generation of markerfree mutations: {{PubMed|20134232,21542786,24282588}}
  
 
==RNA analysis==
 
==RNA analysis==
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<pubmed>23340847 </pubmed>
 
<pubmed>23340847 </pubmed>
 
==Key references==
 
==Key references==
<pubmed>22179594 20057169 20134232 21542786 20300532 19698693 17932079 21841760 17994626 23956387 22285978,22113911 24002656 </pubmed>
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<pubmed>22179594 20057169 20134232 21542786 20300532 19698693 17932079 21841760 17994626 23956387 22285978,22113911 24002656 24282588 </pubmed>

Revision as of 19:05, 28 November 2013

Here you can find a list of methods for working with Bacillus subtilis:

Media

Analysis of protein-protein interactions

  • Bacterial Two Hybrid: BACTH

Analysis of membrane proteins

  • application of a split green fluorescent protein reporter PubMed

Protein expression

Strain construction

  • introduction of markerfree deletions: PubMed
  • generation of markerfree mutations: PubMed

RNA analysis

  • Mechanical cell disruption and extraction of RNA from B. subtilis: Media:SOP-RNA.pdf

Genetic work with non-standard strains

  • transformation of wild-type B. subtilis strains incl. NCIB3610: PubMed
  • efficient electroporation of B. subtilis PubMed
  • a novel transformation protocol for B. subtilis DB104 PubMed

General procedures

  • Standard operation procedures (SOPs) of the SYSMO-BACELL consortium

Microscopic techniques

  • Live cell imaging of B. subtilis cells using automated time-lapse microscopy PubMed
  • Assay of gene expression dynamics using live cell imaging PubMed
  • Visualization and quantification of gene expression heterogeneity in growing microbial cells PubMed
  • GFP variants specifically optimizd for use in B. subtilis PubMed

Transposon mutagenesis

  • mariner transposon mutagenesis for random inducible-expression insertions and transcriptional reporter fusion insertions PubMed

Protein-DNA interactions

  • ChAP-chip: A modified ChIP-chip protocol for the in vivo identification of binding sites of DNA-binding proteins PubMed

Genome engineering

  • generalized bacterial genome editing using mobile group II introns and Cre-lox PubMed
  • A review on genome engineering:

Key references